OBJECTIVE: To identify a candidate neonatal housekeeping gene and to determine the effects of pH and PaO(2) on the stability of newborn gene expression in physiologically hypoxic and acidotic newborn blood. METHODS: Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) amplification was performed for four commonly used housekeeping genes (GAPDH, beta-actin, cyclophilin, 28S rRNA) on extracted RNA. Blood gas analyses determined pH and PaO(2) levels. RESULTS AND CONCLUSIONS: Beta-Actin was the least variable and GAPDH the most variable housekeeping gene studied. pH negatively correlated with gene expression levels. PaO(2) levels did not significantly affect gene expression. These results inform selection of housekeeping genes for neonatal mRNA research.
OBJECTIVE: To identify a candidate neonatal housekeeping gene and to determine the effects of pH and PaO(2) on the stability of newborn gene expression in physiologically hypoxic and acidotic newborn blood. METHODS: Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) amplification was performed for four commonly used housekeeping genes (GAPDH, beta-actin, cyclophilin, 28S rRNA) on extracted RNA. Blood gas analyses determined pH and PaO(2) levels. RESULTS AND CONCLUSIONS:Beta-Actin was the least variable and GAPDH the most variable housekeeping gene studied. pH negatively correlated with gene expression levels. PaO(2) levels did not significantly affect gene expression. These results inform selection of housekeeping genes for neonatal mRNA research.
Authors: Bernd Sehringer; Hans Peter Zahradnik; Wolfgang R Deppert; Michael Simon; Claudia Noethling; Wolfgang R Schaefer Journal: Anal Bioanal Chem Date: 2005-11-05 Impact factor: 4.142
Authors: Enders K O Ng; Nancy B Y Tsui; Nicole Y L Lam; Rossa W K Chiu; Simon C H Yu; S C Cesar Wong; Elena S F Lo; Timothy H Rainer; Philip J Johnson; Y M Dennis Lo Journal: Clin Chem Date: 2002-08 Impact factor: 8.327
Authors: Jo Vandesompele; Katleen De Preter; Filip Pattyn; Bruce Poppe; Nadine Van Roy; Anne De Paepe; Frank Speleman Journal: Genome Biol Date: 2002-06-18 Impact factor: 13.583